Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov 410049, Russia.
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov 410049, Russia.
Talanta. 2019 Sep 1;202:362-368. doi: 10.1016/j.talanta.2019.04.086. Epub 2019 May 4.
A sheep single-chain antibody-fragment library (Griffin.1, UK) was used to obtain miniantibodies to the lipopolysaccharide of Herbaspirillum seropedicae Z78. Using electro-optical analysis and electron microscopy, we recorded a biospecific interaction of antigenic determinants on the cell surface with phage antibodies against the LPS of H. seropedicae Z78 (mini-Abs). Control experiments were run to rule out nonspecific binding of the mini-Abs to cells of Azospirillum brasilense Sp245. Use of the highly specific mini-Abs enabled the lipopolysaccharide of H. seropedicae Z78 to be detected in a mixture of bacterial cells by electro-optical means (analysis time, ∼5 min). This report is the first to show the possibility of rapid detection of Herbaspirillum on the basis of electro-optical analysis coupled with the use of mini-Abs. The results are promising for the development of biosensor-based methods to detect potentially human-harmful prokaryotes whose structures either have not been studied or are absent from commercial databases.
利用绵羊单链抗体片段文库(英国 Griffin.1),我们获得了针对 Herbaspirillum seropedicae Z78 脂多糖的小型抗体。通过光电分析和电子显微镜,我们记录了细胞表面抗原决定簇与针对 H. seropedicae Z78 LPS 的噬菌体抗体(小型 Abs)的生物特异性相互作用。进行了对照实验以排除小型 Abs 与 Azospirillum brasilense Sp245 细胞的非特异性结合。使用高度特异性的小型 Abs 使得能够通过光电手段(分析时间约为 5 分钟)检测混合细菌细胞中的 H. seropedicae Z78 脂多糖。这是首次报道基于光电分析与小型 Abs 结合使用,快速检测 Herbaspirillum 的可能性。这些结果为开发基于生物传感器的方法检测潜在的对人体有害的原核生物提供了希望,这些生物的结构要么尚未研究,要么不在商业数据库中。